The normal hepatocyte LO2 cells, as well as the commercially available HCC cell lines Huh-7, HepG2, HCC-LM3, SMMC-7721 and MHCC-97L, were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Home-made patient-derived HCC cell lines PDC-26#, PDC-14#, PDC-12#, PDC-9#, PDC-23#, PDC-10# and PDC-11# were established as described above. All of these cells were cultured in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% (v/v) FBS and 1% penicillin-streptomycin at 37 °C in a humidified incubator under 5% CO2 condition.
Hcclm3
Manufactured by Shanghai Cell Bank
Sourced in China
The HCCLM3 is a cell culture incubator designed for maintaining and growing cell cultures. It provides a controlled environment with precise temperature, humidity, and CO2 regulation to support the optimal growth and proliferation of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Hepg2, by Shanghai Cell Bank (17 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (15 mentions)
Smmc 7721, by Shanghai Cell Bank (11 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions)
Hep3b, by Shanghai Cell Bank (8 mentions)
Sk hep 1, by Shanghai Cell Bank (7 mentions)
Bel 7402, by Shanghai Cell Bank (7 mentions)
Qgy7701, by Shanghai Cell Bank (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Mhcc 97h, by Shanghai Cell Bank (4 mentions)
Mhcc97l, by Shanghai Cell Bank (3 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Hl 7702, by Shanghai Cell Bank (2 mentions)
Hepg2, by Thermo Fisher Scientific (2 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (2 mentions)
Dmem medium, by Thermo Fisher Scientific (2 mentions)
Hcclm3, by Thermo Fisher Scientific (2 mentions)
Smmc 7721, by Thermo Fisher Scientific (2 mentions)
28 protocols using hcclm3
1
Culturing Human Hepatocellular Carcinoma Cells
2
Culturing Human Liver Cell Lines
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Human HCC cell lines Huh7, Hep3B, HCC-LM3, Sk-Hep-1, and a fetal liver-derived cell line L02 were purchased from Shanghai Cell Bank of the Chinese Academy of Sciences Shanghai, China. All of cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Corning, Inc., Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (M&C Gene Technology Ltd.). Cells were incubated in a thermostatic incubator at 37 °C with 5% CO2.
3
Culturing HCC Cell Lines
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HCC cell lines QGY-7701(Catalogue number:TCHu42), Bel-7402(Catalogue number:TCHu10) and HCCLM3(Catalogue number:TCHu94)were purchased from Shanghai Cell Bank, Chinese Academy of Sciences, MHCC97L cell line(Product number: BNCC337741)was purchased from Bei Na Chuanglian Institute of Biotechnology of Beijing, china. All cell lines were cultured in DMEM or RPMI-1640 medium, which contained 10% FBS (Gibco, USA). As described previously, cells were cultured at 37 °C under 5% CO2 [4 (link)].
4
Culturing Human Liver Cancer Cell Lines
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Human HCC cell lines HepG2, Hep3b, BEL7404, Huh7, SKhep1, and PLCPRF5 were all purchased from American Type Culture Collection (ATCC, Manassas, VA). Human normal liver cell line HL7702 and human HCC cell lines BEL7402, BEL7405, HCCLM3, MHCC97h, QGY-7701, QGY-7703, and SMMC-7721 were purchased from Cell Bank of Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). All cells were cultured in Dulbecco modified Eagle medium (Thermo Fisher Scientific Inc, Waltham, MA) containing 10% fetal bovine serum (Thermo Fisher Scientific, Inc), 1% penicillin and streptomycin solution at 37°C in an incubator with a humidified atmosphere and 5% CO2.
5
Cohort Study of HCC Tissue and Cells
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Two independent cohorts composed of 216 patients with HCC were enrolled in this study (cohort 1, n = 136; cohort 2, n = 80). Paired HCC tissues and adjacent liver tissues were collected from patients who underwent liver resection at the First Affiliated Hospital of Harbin Medical University between January 2008 and August 2013. All patients provided written informed consent and this study was approved by the Research Ethics Committee of the First Affiliated Hospital of Harbin Medical University. Follow-up data were collected from patients after hepatic resection to monitor and assess the overall rate of cancer metastasis and recurrence. A normal liver cell line (L02) and several HCC cell lines (Huh7, HCCLM3, SK-Hep-1, MHCC97H, MHCC97L, SMMC7721, and HepG2) were purchased from Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China).
6
Liver Tumor Tissue Acquisition and Cell Culture
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We obtained 49 pairs of HCC and adjacent liver tissues from individuals that had undergone liver tumor resection at the First Affiliated Hospital of Harbin Medical University from January 2016 to January 2018. All participants were asked to sign an informed consent form. The research ethics committee of the hospital approved the study. There were 5 cell lines utilized in this study, including a normal liver cell line (L02) and four HCC cell lines (SK-Hep-1, Huh7, HCCLM3 and HepG2) that were bought from Shanghai Cell Bank of Chinese Academy of Sciences. All human HCC cell lines were grown in DMEM complete medium culture plus 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin and incubated at 37 °C with 5% CO2.
7
Cell Culture of Liver Cell Lines
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L02, HepG2, HCCLM3, MHCC97H, and BEL-7702 cell lines were purchased from Shanghai Cell Bank of the Chinese Academy of Science (Shanghai, People’s Republic of China). All cell lines were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS; Gibco), and then incubated with 5% CO2 at 37°C in a humidified atmosphere.
8
Evaluation of ATIC in Liver Cancer Cells
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Huh7, HepG2 and HCCLM3 liver cancer cells were obtained from Shanghai Cell Bank (Shanghai, China). The cell lines had recently been authenticated and tested for mycoplasma. Huh7, HepG2 and HCCLM3 cells were grown in DMEM (Invitrogen, Carlsbad, CA). All media were supplemented with 10% fetal bovine serum (Invitrogen). Jtsbio (Guangzhou, China) constructed and synthesized the siRNA and shRNA lentivirus vector specific for ATIC. The sequence of the shRNA is the same as siRNA 3:
ATIC-siRNA 1: forward, 5'-CCUGCAAUCUCUAUCCCUUTT -3'; reverse, 5'-AAGGGAUAGAGAUUGCAGGTT -3' and;
ATIC-siRNA 3: forward, 5'-GGUGUCGUCGACAAGUCAUTT -3'; reverse, 5'-AUGACUUGUCGACGACACCTT -3'.
For ATIC overexpression, the full-length ATIC cDNA was amplified by PCR and cloned into the expression vector pcDNA3.1 (Invitrogen). The empty vector was used as a control. Transfection was performed using Lipofectamine 3000 reagent (Invitrogen). Forty-eight hours after transfection, cells were harvested for further analysis.
ATIC-siRNA 1: forward, 5'-CCUGCAAUCUCUAUCCCUUTT -3'; reverse, 5'-AAGGGAUAGAGAUUGCAGGTT -3' and;
ATIC-siRNA 3: forward, 5'-GGUGUCGUCGACAAGUCAUTT -3'; reverse, 5'-AUGACUUGUCGACGACACCTT -3'.
For ATIC overexpression, the full-length ATIC cDNA was amplified by PCR and cloned into the expression vector pcDNA3.1 (Invitrogen). The empty vector was used as a control. Transfection was performed using Lipofectamine 3000 reagent (Invitrogen). Forty-eight hours after transfection, cells were harvested for further analysis.
9
Characterization of Liver Cancer Cell Lines
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Human liver cancer cells (HCC-LM3, Huh7, Hep-G2, Hep-G2.215, SK-Hep1 and 7721) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. The human liver cell line HL-7702 (L02) was purchased from Wuhan Hengyisai Biotechnology Co., Ltd. Short tandem repeat (STR) profiling was used to identify all cells. All cells were cultured in an incubator at 37°C and 5% carbon dioxide using DMEM with 10% FBS.
10
Liver Cancer Tissue Collection and Cell Culture
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Eighty HCC tissues and corresponding noncancerous adjoining tissues (NATs) were harvested from HCC patients who received surgical intervention at the First Affiliated Hospital of China Medical University (Shenyang, China) between 2018 and 2019. All participants were pathologically confirmed post‐surgery, and signed an informed consent form. NATs were extracted from regions over 2 cm away from the tumor. All tissue samples were quickly flash frozen in liquid nitrogen following harvest, and before transfer to −80°C for prolonged storage. This work received ethical approval from our institution. All liver cancer cells, namely, Bel7402, Huh7, HCCLM3, SMMC7721, HepG2, and LO2, were acquired from the Shanghai Cell Bank (Shanghai, China). All cells were grown in high‐glucose Dulbecco's‐modified eagle medium (HyClone) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37°C in a 5% CO2 incubator (Thermo).
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